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Interaction studies of a protein and carbohydrate system using an integrated approach: a case study of the miniagrin–heparin system

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Abstract

The major challenges in biophysical characterization of human protein–carbohydrate interactions are obtaining monodispersed preparations of human proteins that are often post-translationally modified and lack of detection of carbohydrates by traditional detection systems. Light scattering (dynamic and static) techniques offer detection of biomolecules and their complexes based on their size and shape, and do not rely on chromophore groups (such as aromatic amino acid sidechains). In this study, we utilized dynamic light scattering, analytical ultracentrifugation and small-angle X-ray scattering techniques to investigate the solution properties of a complex resulting from the interaction between a 15 kDa heparin preparation and miniagrin, a miniaturized version of agrin. Results from dynamic light scattering, sedimentation equilibrium, and sedimentation velocity experiments signify the formation of a monodisperse complex with 1:1 stoichiometry, and low-resolution structures derived from the small-angle X-ray scattering measurements implicate an extended conformation for a side-by-side miniagrin‒heparin complex.

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Acknowledgements

TRP thanks the Canadian Institutes of Health Research for PDF. He is a Canada Research Chair in RNA and Protein Biophysics. JS holds the Canada Research Chair in Structural Biology and Biophysics. Professor Markus Ruegg kindly provided the minagrin gene.

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Authors and Affiliations

  1. Department of Chemistry and Biochemistry, Alberta RNA Research and Training Institute, University of Lethbridge, 4401 University Drive, Lethbridge, AB, T1K 3M4, Canada

    Trushar R. Patel

  2. Discovery Lab, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, AB, T6G 2H7, Canada

    Trushar R. Patel

  3. Department of Microbiology, Immunology and Infectious Diseases, Cumming School of Medicine, University of Calgary, 2500 University Dr. NW, Calgary, AB, T2N 1N4, Canada

    Trushar R. Patel

  4. National Centre for Macromolecular Hydrodynamics, University of Nottingham, Sutton Bonington, LE12 5RsD, UK

    Tabot M. D. Besong & Stephen E. Harding

  5. Department of Chemistry, University of Manitoba, Winnipeg, MB, R3T 2N2, Canada

    Markus Meier, Kevin McEleney & Jörg Stetefeld

  6. Manitoba Institute for Materials, University of Manitoba, Winnipeg, MB, R3T 2N2, Canada

    Kevin McEleney

  7. School of Chemistry and Molecular Biosciences, University of Queensland, Brisbane, QLD, 4072, Australia

    Donald J. Winzor

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  1. Trushar R. Patel
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  2. Tabot M. D. Besong
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  3. Markus Meier
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  4. Kevin McEleney
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  5. Stephen E. Harding
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  7. Jörg Stetefeld
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Correspondence to Trushar R. Patel or Jörg Stetefeld.

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Special Issue: 23rd International AUC Workshop and Symposium.

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Patel, T.R., Besong, T.M.D., Meier, M. et al. Interaction studies of a protein and carbohydrate system using an integrated approach: a case study of the miniagrin–heparin system. Eur Biophys J 47, 751–759 (2018). https://doi.org/10.1007/s00249-018-1291-5

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  • DOI: https://doi.org/10.1007/s00249-018-1291-5

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